Regeneration of multiple shoots from transgenic potato events facilitates the recovery of phenotypically normal lines: assessing a cry9Aa2 gene conferring insect resistance

Background

Tools for authenticating cell lines are critical for quality control in cell-based biological experiments. Currently there are methods to authenticate human cell lines using short tandem repeat (STR) markers based on the technology and procedures successfully used in the forensic community for human identification, but there are no STR based methods for authenticating nonhuman cell lines to date. There is significant homology between the human and vervet monkey genome and we utilized these similarities to design the first multiplex assay based on human STR markers for vervet cell line identification.

Results

The following STR markers were incorporated into the vervet multiplex PCR assay: D17S1304, D5S1467, D19S245, D1S518, D8S1106, D4S2408, D6S1017, and DYS389. The eight markers were successful in uniquely identifying sixty-two vervet monkey DNA samples and confirmed that Vero76 cells and COS-7 cells were derived from Vero and CV-1 cells, respectively. The multiplex assay shows specificity for vervet DNA within the determined allele range for vervet monkeys; however, the primers will also amplify human DNA for each marker resulting in amplicons outside the vervet allele range in several of the loci. The STR markers showed genetic stability in over sixty-nine passages of Vero cells, suggesting low mutation rates in the targeted STR sequences in the Vero cell line.

Conclusions

A functional vervet multiplex assay consisting of eight human STR markers with heterozygosity values ranging from 0.53-0.79 was successful in uniquely identifying sixty-two vervet monkey samples. The probability of a random match using these eight markers between any two vervet samples is approximately 1 in 1.9 million. While authenticating a vervet cell line, the multiplex assay may also be a useful indicator for human cell line contamination since the assay is based on human STR markers.     

The plant ant Tetraponera aethiops (Pseudomyrmecinae) protects its host myrmecophyte Barteria fistulosa (Passifloraceae) through aggressiveness and predation

Plant ants generally provide their host myrmecophytes (i.e. plants that shelter a limited number of ant species in hollow structures) protection from defoliating insects, but the exact nature of this protection is poorly known. It was with this in mind that we studied the association between Tetraponera aethiops F. Smith (Pseudomyrmecinae) and its specific host myrmecophyte Barteria fistulosa Mast. (Passifloraceae). Workers bore entrances into the horizontal hollow branches (domatia) of their host B. fistulosa , near the base of the petiole of the alternate horizontal leaves. They then ambush intruders from the domatia, close to these entrances. After perceiving the vibrations caused when an insect lands on a leaf, they rush to it and sting and generally spreadeagle the insect (only small caterpillars are mastered by single workers). Among the insects likely to defoliate B. fistulosa , adult leaf beetles and large katydids were taken as prey and cut up; single workers then retrieved some pieces, whereas other workers imbibed the prey's haemolymph. Other insects known to defoliate this plant, if unable to escape, were killed and discarded. Small Acrea zetes L. caterpillars were stung and then discarded by single workers; whereas locusts of different sizes were mastered by groups of workers that stung and spreadeagled them before discarding them (although a part of their haemolymph was imbibed). More workers were involved and more time was necessary to master insects taken as prey than those attacked and discarded. Consequently, the protection T. aethiops workers provide to their host B. fistulosa from defoliating insects results from predation, but more often from a type of aggressiveness wherein insects are killed and then discarded.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 93 , 63–69.     

Simultaneous saccharification and fermentation of pretreated biomass: Improving mass balance closure

Summary The successful implementation of simultaneous saccharification and fermentation (SSF) technology for biomass conversion to ethanol requires competent analysis of complex biomass process streams, often obtained at extremes of pH. In this study, optimal conditions for handling biomass samples recovered from acid and alkaline pretreatments prior to traditional compositional analysis were developed. Methods for processing slurries from SSF were also determined. In both cases, a mixed wastepaper feedstock was used to test improved handling procedures and to document recommended performance.     

Parasitoid vibrations as potential releasing stimulus of evasive behaviour in a leafminer

Abstract. The aim of this study was to characterize the vibratory signals produced by the parasitoid Sympiesis sericeicornis Nees (Hymenoptera: Eulophidae) while foraging on apple leaves infested by one of its hosts, the spotted tentiform leafminer Phyllonorycter malella (Ger.) (Lepidoptera: Gracillariidae).This leafminer changes its behaviour as a function of the parasitoid's behaviour to escape parasitization.We propose that the leafminer uses vibrations triggered by the parasitoid to detect the presence of its enemy.We measured vibrations produced by a foraging parasitoid on a mine with a laser vibrometer.By recording concurrently the behaviour of the parasitoid on video, vibrations could be assigned to particular behaviours.Subsequently, vibrations were characterized by their dominant frequencies and intensities.The behaviours Landing and Take-off both produced strong impact-like vibrations characterized by an initial irregular phase during which frequencies up to 25 kHz occurred followed by a slow decaying regular phase.Vibrations elicited by Moving, Standing and Probing showed no clear temporal pattern.During Probing , dominant frequencies of up to 5.6 kHz were observed frequently at intensities well above the background noise (>10 dB).During Moving and Standing , vibrations were more scarce and of lower frequencies and intensities.Due to their impact-like nature, vibrations produced by Landing and Take-off are probably not specific to the parasitoid.Vibrations produced by Moving and Standing are difficult to detect and not reliable because of their non-specificity.Therefore, only Probing provides a reliable and detectable source of information for the host.The vibrations elicited during Probing could account for the evasive behaviour that is observed in this and other leafminers.